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  • 95
    Thermo Fisher igg4 human elisa kit
    Immunofluorescence and <t>ELISA-based</t> assessment of antibody secretion. A: Brightfield image of a single well, seeded with antibody-secreting B13-24 CHO cells. Aggregates of secreted antibodies are annotated for visualization using white pixels. High-producing colonies are annotated using red rectangles. B-E: Enlarged annotated bright field (B,D) and fluorescence (C,E) images of high and low producing colonies. F: Secreted <t>IgG4</t> concentration as measured by ELISA of supernatant sampled from the 96-well plate on day 15 (N = 5). G: High-producer cells maintained their increased IgG4 production during expansion up to 25 days into T-25 culture flasks, compared to control and low-producers. (Scale bar = 50 μm).
    Igg4 Human Elisa Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc horseradish peroxidase conjugated anti rabbit immunoglobulin igg
    Immunofluorescence and <t>ELISA-based</t> assessment of antibody secretion. A: Brightfield image of a single well, seeded with antibody-secreting B13-24 CHO cells. Aggregates of secreted antibodies are annotated for visualization using white pixels. High-producing colonies are annotated using red rectangles. B-E: Enlarged annotated bright field (B,D) and fluorescence (C,E) images of high and low producing colonies. F: Secreted <t>IgG4</t> concentration as measured by ELISA of supernatant sampled from the 96-well plate on day 15 (N = 5). G: High-producer cells maintained their increased IgG4 production during expansion up to 25 days into T-25 culture flasks, compared to control and low-producers. (Scale bar = 50 μm).
    Horseradish Peroxidase Conjugated Anti Rabbit Immunoglobulin Igg, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 2857 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    EUROIMMUN igg elisa
    Anti-ZIKV reactivity in potentially cross-reactive samples (n = 252) and healthy controls (n = 1,015) as determined by <t>ELISA</t> for (A) IgM and (B) <t>IgG</t> a,b , study evaluating a novel NS1-based ELISA, Germany 2016*
    Igg Elisa, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 90/100, based on 66 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    EUROIMMUN anti vzv elisa igg
    Comparison of quantitative results obtained with the Enzygnost <t>antiVZV/IgG,</t> Serion <t>ELISA</t> Classic <t>VZV</t> IgG, and Euroimmun anti-VZV ELISA (IgG) tests for positive sera from blood donors of panel 2. A total of 357 sera within the upper quantification limit
    Anti Vzv Elisa Igg, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 88/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    EUROIMMUN anti sars cov 2 elisa igg
    Relation of Ct values of the RT-PCR runs and <t>Anti-SARS-CoV-2-ELISA</t> IgG ratio in symptomatic outpatients (A) and asymptomatic contact persons (B). Figure legend: Ct values of 94 outpatients and 23 asymptomatic contact persons were plotted against SARS-CoV-2-IgG ratios determined by SARS-CoV-2-ELISA IgG (Euroimmun). Data included ct values for the E gene (n = 82 patients and n = 19 contact persons), orf gene (n = 50 patients and n = 8 contact persons), and N gene (n = 20 patients and N = 2 contact persons).
    Anti Sars Cov 2 Elisa Igg, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    EUROIMMUN anti zika virus elisa igg
    (a–e) Show the relationship between days post-onset of symptoms (DPO) and kit results for samples that tested ZIKV IgM-positive using reference methods. Numbers of samples tested at each time point are listed on the y-axis. Results in Fig. 1a for InBios ZIKV Detect™ IgM Capture <t>ELISA</t> were generated using serum Panels 1 and 2; results in Fig. 1b. for NovaTec NovaLisa® <t>Zika</t> Virus IgM μ-capture ELISA were generated using Panels 1, 2 and 3, where equivocals were classified as Zika Positive; results for Euroimmun Anti-Zika Virus ELISA IgM and <t>IgG</t> (Fig. 1c-e) were generated using Panel 1, where borderlines were classified as Zika Positive.
    Anti Zika Virus Elisa Igg, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Alpha Diagnostics mouse anti dsdna igg elisa kit
    Forced expression of Igβ in B cells impaired the antibody production and affinity maturation. ( a ) Expression of Igβ in B220 + CD38 − GL7 − splenocytes from immunized Igβ-Tg (Igβ-Tg; line) and wild-type litter mates (wild type; shaded). ( b ) Expression of CD38 and GL7 in B220 + spleen cells from immunized wild type litter mates ( top left ) and Igβ-Tg mice ( top right ). Cell cycle analysis of CD38 − GL7 − GC B cells from wild type litter mates ( bottom left ) and Igβ-Tg mice ( bottom right ). Numbers indicate the percentages of cells in each gate. ( c ) Serum anti-NP titers in each isotype after the immunization. Igβ-Tg (open circle) and wild type litter mates (closed circle) were immunized with NP-CGG on day 0 (1°) and boosted 42 d later (2°). Sera were collected at indicated day and assayed by <t>ELISA</t> as described in Methods section. Error bars indicate SEM for each group. Each group consists of 5 mice. Representative data from 3 repeated experiments is shown. ( d ) Affinity maturation of anti-NP <t>IgG1</t> antibody in NP-CGG immunized Igβ-Tg (closed bar) and wild type litter mates (open bar). Error bars indicate SEM for each group. ( e ) Decreased frequency of somatic hypermutation in immunized Igβ-Tg mice. CD38 − GL7 + GC B cells were isolated from Igβ-Tg and wild type litter mates 21 days after the immunization. Total RNA was extracted and reverse transcribed to obtain cDNA samples. VH186.2 gene was then amplified and sequenced for the mutation analysis.
    Mouse Anti Dsdna Igg Elisa Kit, supplied by Alpha Diagnostics, used in various techniques. Bioz Stars score: 88/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Focus Diagnostics Inc herpeselect 2 elisa igg kit
    Forced expression of Igβ in B cells impaired the antibody production and affinity maturation. ( a ) Expression of Igβ in B220 + CD38 − GL7 − splenocytes from immunized Igβ-Tg (Igβ-Tg; line) and wild-type litter mates (wild type; shaded). ( b ) Expression of CD38 and GL7 in B220 + spleen cells from immunized wild type litter mates ( top left ) and Igβ-Tg mice ( top right ). Cell cycle analysis of CD38 − GL7 − GC B cells from wild type litter mates ( bottom left ) and Igβ-Tg mice ( bottom right ). Numbers indicate the percentages of cells in each gate. ( c ) Serum anti-NP titers in each isotype after the immunization. Igβ-Tg (open circle) and wild type litter mates (closed circle) were immunized with NP-CGG on day 0 (1°) and boosted 42 d later (2°). Sera were collected at indicated day and assayed by <t>ELISA</t> as described in Methods section. Error bars indicate SEM for each group. Each group consists of 5 mice. Representative data from 3 repeated experiments is shown. ( d ) Affinity maturation of anti-NP <t>IgG1</t> antibody in NP-CGG immunized Igβ-Tg (closed bar) and wild type litter mates (open bar). Error bars indicate SEM for each group. ( e ) Decreased frequency of somatic hypermutation in immunized Igβ-Tg mice. CD38 − GL7 + GC B cells were isolated from Igβ-Tg and wild type litter mates 21 days after the immunization. Total RNA was extracted and reverse transcribed to obtain cDNA samples. VH186.2 gene was then amplified and sequenced for the mutation analysis.
    Herpeselect 2 Elisa Igg Kit, supplied by Focus Diagnostics Inc, used in various techniques. Bioz Stars score: 88/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    GenWay anti c trachomatis igg elisas
    Detection sensitivity of anti- C. <t>trachomatis</t> antibodies in sera from Ctr-NAAT-positive women. An artificial population was created by combining 125 high anti- C. trachomatis antibody-prevalence sera from Ctr-NAAT-positive women ( 21 ) and 87 sera that had tested negative in multiple previous Ctr antibody assays ( Fig. 1 ). The first (leftmost) column indicates origin of sera from either the cohort of 125 Ctr NAAT-positive women (red) or the 3 cohorts of low Ctr-antibody-prevalence sera shown in Fig. 1 . The next column shows the ethnicity of the subjects as in Fig. 1 , followed by the composite reference standard CRS1 for NAAT-proven exposure to C. trachomatis ( Table 2 ). The next columns show background-corrected colorimetric signals of Ctr Mix1 and Ctr Mix 2 for an anti-human <t>IgG</t> polyclonal antibody conjugate, as well as monoclonal antibody conjugates for detection of short-lived IgG3 and IgA1 A2 antibody isotypes. The cutoff in these assays was chosen at 98.9% or higher specificity, with negative (white) and percentile reactivity ranking from low (light brown) to maximal (dark brown). The next columns indicate reactivity in 4 anti- C. trachomatis IgG commercial <t>ELISAs</t> as partially shown in Fig. 1 , followed by a Chlamydia genus-specific ELISA for detection of IgG antibodies against chlamydial LPS. The last column shows a consensus antibody status (positive/negative) based on both 4 commercial ELISAs and 2 peptide assays for anti- C. trachomatis IgG, IgG3, and IgA antibodies. This consensus status, CRS2, was used for ROC analyses that evaluated the performance of the individual assays ( Table 3 ).
    Anti C Trachomatis Igg Elisas, supplied by GenWay, used in various techniques. Bioz Stars score: 93/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher human igg subclass profile elisa kit
    FOXP1 overexpression specifically inhibits formation of <t>IgG-</t> but not IgM-secreting PCs. (A-B) CD19 + CD27 + MBCs were sorted from human peripheral blood and transduced with FOXP1-IRES-YFP, BCL6-IRES-GFP, or control empty vector and cultured under conditions that promote PC differentiation. Six days after transduction, YFP- or GFP-positive cells of cultures transduced with FOXP1, BCL6, or control vector were sorted. (A) Equal numbers of sorted cells (50 000) were cultured with IL-21 and IL-2 for an additional 24 hours. Thereafter, the supernatants were collected, and IgM and IgG protein levels were analyzed by <t>ELISA.</t> Levels were normalized to levels in control-transduced cells. Means ± SD of at least 4 independent experiments are shown. (B) Equal numbers of sorted cells were plated onto membranes in serial dilutions and cultured with IL-21 and IL-2 for an additional 18 hours, after which numbers of IgM- or IgG-secreting cells were determined by ELISPOT. Spot numbers were normalized to numbers in stimulated, control-transduced cells. Means ± SD of 3 independent experiments are shown (1 sample t test, * P
    Human Igg Subclass Profile Elisa Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Immunofluorescence and ELISA-based assessment of antibody secretion. A: Brightfield image of a single well, seeded with antibody-secreting B13-24 CHO cells. Aggregates of secreted antibodies are annotated for visualization using white pixels. High-producing colonies are annotated using red rectangles. B-E: Enlarged annotated bright field (B,D) and fluorescence (C,E) images of high and low producing colonies. F: Secreted IgG4 concentration as measured by ELISA of supernatant sampled from the 96-well plate on day 15 (N = 5). G: High-producer cells maintained their increased IgG4 production during expansion up to 25 days into T-25 culture flasks, compared to control and low-producers. (Scale bar = 50 μm).

    Journal: bioRxiv

    Article Title: Selective Cell Propagation via Micropatterning of Thermal-activated Hydrogel

    doi: 10.1101/2020.04.01.019208

    Figure Lengend Snippet: Immunofluorescence and ELISA-based assessment of antibody secretion. A: Brightfield image of a single well, seeded with antibody-secreting B13-24 CHO cells. Aggregates of secreted antibodies are annotated for visualization using white pixels. High-producing colonies are annotated using red rectangles. B-E: Enlarged annotated bright field (B,D) and fluorescence (C,E) images of high and low producing colonies. F: Secreted IgG4 concentration as measured by ELISA of supernatant sampled from the 96-well plate on day 15 (N = 5). G: High-producer cells maintained their increased IgG4 production during expansion up to 25 days into T-25 culture flasks, compared to control and low-producers. (Scale bar = 50 μm).

    Article Snippet: B13-24 cells are capable of producing up to 100 μg/L of humanized immunoglobulin IgG4 at a rate of 25 pg/cell/day .

    Techniques: Immunofluorescence, Enzyme-linked Immunosorbent Assay, Fluorescence, Concentration Assay

    Process to select for high-producing CHO cell clones using SLG. A: CHO cells are mixed into a media solution containing 1% methylcellulose and 1% CloneDetect to detect secreted IgG4. B: The suspension is plated into a 96-well plate. C: Each well is scanned in bright field and fluorescence. D: Colonies are ranked based on antibody secretion per cell. E: The top-producing cells were retained, while SLG encapsulates the remaining cells in hydrogel to arrest their growth. F: Liquid media is added and the remaining cells are allowed to propagate. G: Antibody secretion rate can be measured via ELISA by sampling the supernatant. H: The culture is expanded using large containers.

    Journal: bioRxiv

    Article Title: Selective Cell Propagation via Micropatterning of Thermal-activated Hydrogel

    doi: 10.1101/2020.04.01.019208

    Figure Lengend Snippet: Process to select for high-producing CHO cell clones using SLG. A: CHO cells are mixed into a media solution containing 1% methylcellulose and 1% CloneDetect to detect secreted IgG4. B: The suspension is plated into a 96-well plate. C: Each well is scanned in bright field and fluorescence. D: Colonies are ranked based on antibody secretion per cell. E: The top-producing cells were retained, while SLG encapsulates the remaining cells in hydrogel to arrest their growth. F: Liquid media is added and the remaining cells are allowed to propagate. G: Antibody secretion rate can be measured via ELISA by sampling the supernatant. H: The culture is expanded using large containers.

    Article Snippet: B13-24 cells are capable of producing up to 100 μg/L of humanized immunoglobulin IgG4 at a rate of 25 pg/cell/day .

    Techniques: Clone Assay, Fluorescence, Enzyme-linked Immunosorbent Assay, Sampling

    Anti-ZIKV reactivity in potentially cross-reactive samples (n = 252) and healthy controls (n = 1,015) as determined by ELISA for (A) IgM and (B) IgG a,b , study evaluating a novel NS1-based ELISA, Germany 2016*

    Journal: Eurosurveillance

    Article Title: Serodiagnosis of Zika virus (ZIKV) infections by a novel NS1-based ELISA devoid of cross-reactivity with dengue virus antibodies: a multicohort study of assay performance, 2015 to 2016

    doi: 10.2807/1560-7917.ES.2016.21.50.30426

    Figure Lengend Snippet: Anti-ZIKV reactivity in potentially cross-reactive samples (n = 252) and healthy controls (n = 1,015) as determined by ELISA for (A) IgM and (B) IgG a,b , study evaluating a novel NS1-based ELISA, Germany 2016*

    Article Snippet: Enzyme-linked immunosorbent assays Anti-Zika Virus IgM and IgG ELISA (EUROIMMUN) were used as recommended by the manufacturer.

    Techniques: Enzyme-linked Immunosorbent Assay

    Anti-ZIKV reactivity in patients with RT-PCR-confirmed (n = 27) and suspected (n = 85) ZIKV infection as determined by ELISA for (A) IgM and (B) IgG a ; time course analysis of anti-ZIKV antibody levels in follow-up samples from (C) a German patient returning from Colombia (probable primary ZIKV infection) b and (D) a Colombian patient with RT-PCR-confirmed ZIKV infection (probable secondary flavivirus infection) c

    Journal: Eurosurveillance

    Article Title: Serodiagnosis of Zika virus (ZIKV) infections by a novel NS1-based ELISA devoid of cross-reactivity with dengue virus antibodies: a multicohort study of assay performance, 2015 to 2016

    doi: 10.2807/1560-7917.ES.2016.21.50.30426

    Figure Lengend Snippet: Anti-ZIKV reactivity in patients with RT-PCR-confirmed (n = 27) and suspected (n = 85) ZIKV infection as determined by ELISA for (A) IgM and (B) IgG a ; time course analysis of anti-ZIKV antibody levels in follow-up samples from (C) a German patient returning from Colombia (probable primary ZIKV infection) b and (D) a Colombian patient with RT-PCR-confirmed ZIKV infection (probable secondary flavivirus infection) c

    Article Snippet: Enzyme-linked immunosorbent assays Anti-Zika Virus IgM and IgG ELISA (EUROIMMUN) were used as recommended by the manufacturer.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Infection, Enzyme-linked Immunosorbent Assay

    Development of anti-SARS-CoV-2-IgG antibody response over time in COVID-19 patients and COVID-19-contact groups. Shown is the number of positive results in the Euroimmun IgG-testing at time points “baseline” (dark) and “follow up” (light) relative to all tests per group at the given time. The results are displayed separately for health care workers with initially positive PCR testing for SARS-CoV-2 (“COVID-19 patients”) and health care workers with negative PCR (“COVID-19 contact groups”). * p

    Journal: Journal of Clinical Virology

    Article Title: Immune response to SARS-CoV-2 in health care workers following a COVID-19 outbreak: A prospective longitudinal study

    doi: 10.1016/j.jcv.2020.104575

    Figure Lengend Snippet: Development of anti-SARS-CoV-2-IgG antibody response over time in COVID-19 patients and COVID-19-contact groups. Shown is the number of positive results in the Euroimmun IgG-testing at time points “baseline” (dark) and “follow up” (light) relative to all tests per group at the given time. The results are displayed separately for health care workers with initially positive PCR testing for SARS-CoV-2 (“COVID-19 patients”) and health care workers with negative PCR (“COVID-19 contact groups”). * p

    Article Snippet: Among the COVID-19 cases, the variability of values was wide: IgG values ranged from 0.1 to 7.3 using the Euroimmun IgG ELISA and the antibody values from the Elecsys ranged from 0.1 to 128.3.

    Techniques: Polymerase Chain Reaction

    Comparison of quantitative results obtained with the Enzygnost antiVZV/IgG, Serion ELISA Classic VZV IgG, and Euroimmun anti-VZV ELISA (IgG) tests for positive sera from blood donors of panel 2. A total of 357 sera within the upper quantification limit

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Evaluation of Three Commercial Varicella-Zoster Virus IgG Enzyme-Linked Immunosorbent Assays in Comparison to the Fluorescent-Antibody-to-Membrane-Antigen Test

    doi: 10.1128/CVI.00183-12

    Figure Lengend Snippet: Comparison of quantitative results obtained with the Enzygnost antiVZV/IgG, Serion ELISA Classic VZV IgG, and Euroimmun anti-VZV ELISA (IgG) tests for positive sera from blood donors of panel 2. A total of 357 sera within the upper quantification limit

    Article Snippet: Correlation with the reference assay (FAMA) was computed for the Enzygnost anti-VZV/IgG, Serion ELISA Classic VZV IgG, and Euroimmun anti-VZV ELISA (IgG) assays as the number of sera with concordant results divided by the total number of serum samples included.

    Techniques: Enzyme-linked Immunosorbent Assay

    Comparison of quantitative results obtained by FAMA with those of the Enzygnost anti-VZV/IgG, Serion ELISA Classic VZV IgG, and Euroimmun anti-VZV ELISA (IgG) tests with 419 positive sera from the blood donors of panel 2. Each dot is the results of a

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Evaluation of Three Commercial Varicella-Zoster Virus IgG Enzyme-Linked Immunosorbent Assays in Comparison to the Fluorescent-Antibody-to-Membrane-Antigen Test

    doi: 10.1128/CVI.00183-12

    Figure Lengend Snippet: Comparison of quantitative results obtained by FAMA with those of the Enzygnost anti-VZV/IgG, Serion ELISA Classic VZV IgG, and Euroimmun anti-VZV ELISA (IgG) tests with 419 positive sera from the blood donors of panel 2. Each dot is the results of a

    Article Snippet: Correlation with the reference assay (FAMA) was computed for the Enzygnost anti-VZV/IgG, Serion ELISA Classic VZV IgG, and Euroimmun anti-VZV ELISA (IgG) assays as the number of sera with concordant results divided by the total number of serum samples included.

    Techniques: Enzyme-linked Immunosorbent Assay

    Relation of Ct values of the RT-PCR runs and Anti-SARS-CoV-2-ELISA IgG ratio in symptomatic outpatients (A) and asymptomatic contact persons (B). Figure legend: Ct values of 94 outpatients and 23 asymptomatic contact persons were plotted against SARS-CoV-2-IgG ratios determined by SARS-CoV-2-ELISA IgG (Euroimmun). Data included ct values for the E gene (n = 82 patients and n = 19 contact persons), orf gene (n = 50 patients and n = 8 contact persons), and N gene (n = 20 patients and N = 2 contact persons).

    Journal: Journal of Clinical Virology

    Article Title: SARS-CoV-2-IgG response is different in COVID-19 outpatients and asymptomatic contact persons

    doi: 10.1016/j.jcv.2020.104542

    Figure Lengend Snippet: Relation of Ct values of the RT-PCR runs and Anti-SARS-CoV-2-ELISA IgG ratio in symptomatic outpatients (A) and asymptomatic contact persons (B). Figure legend: Ct values of 94 outpatients and 23 asymptomatic contact persons were plotted against SARS-CoV-2-IgG ratios determined by SARS-CoV-2-ELISA IgG (Euroimmun). Data included ct values for the E gene (n = 82 patients and n = 19 contact persons), orf gene (n = 50 patients and n = 8 contact persons), and N gene (n = 20 patients and N = 2 contact persons).

    Article Snippet: 2.1.1 Immunoassay for SARS-CoV-2 antibody testingSARS-CoV-2-IgG antibodies were determined within three days after receipt of samples by Anti-SARS-CoV-2-ELISA IgG (Euroimmun, Luebeck, Germany, antigen S1 spike protein) on the Euroimmun Workstation ELISA according to the manufacturer’s instructions.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    (a–e) Show the relationship between days post-onset of symptoms (DPO) and kit results for samples that tested ZIKV IgM-positive using reference methods. Numbers of samples tested at each time point are listed on the y-axis. Results in Fig. 1a for InBios ZIKV Detect™ IgM Capture ELISA were generated using serum Panels 1 and 2; results in Fig. 1b. for NovaTec NovaLisa® Zika Virus IgM μ-capture ELISA were generated using Panels 1, 2 and 3, where equivocals were classified as Zika Positive; results for Euroimmun Anti-Zika Virus ELISA IgM and IgG (Fig. 1c-e) were generated using Panel 1, where borderlines were classified as Zika Positive.

    Journal: Journal of virological methods

    Article Title: Multi-laboratory comparison of three commercially available Zika IgM enzyme-linked immunosorbent assays

    doi: 10.1016/j.jviromet.2018.06.018

    Figure Lengend Snippet: (a–e) Show the relationship between days post-onset of symptoms (DPO) and kit results for samples that tested ZIKV IgM-positive using reference methods. Numbers of samples tested at each time point are listed on the y-axis. Results in Fig. 1a for InBios ZIKV Detect™ IgM Capture ELISA were generated using serum Panels 1 and 2; results in Fig. 1b. for NovaTec NovaLisa® Zika Virus IgM μ-capture ELISA were generated using Panels 1, 2 and 3, where equivocals were classified as Zika Positive; results for Euroimmun Anti-Zika Virus ELISA IgM and IgG (Fig. 1c-e) were generated using Panel 1, where borderlines were classified as Zika Positive.

    Article Snippet: 3) The combined assays Anti-Zika Virus ELISA (IgM) and Anti-Zika Virus ELISA (IgG) (Euroimmun Medizinische Labordiagnostika AG (Lübeck, Germany), both use ZIKV NS1 antigen.

    Techniques: Enzyme-linked Immunosorbent Assay, Generated

    Forced expression of Igβ in B cells impaired the antibody production and affinity maturation. ( a ) Expression of Igβ in B220 + CD38 − GL7 − splenocytes from immunized Igβ-Tg (Igβ-Tg; line) and wild-type litter mates (wild type; shaded). ( b ) Expression of CD38 and GL7 in B220 + spleen cells from immunized wild type litter mates ( top left ) and Igβ-Tg mice ( top right ). Cell cycle analysis of CD38 − GL7 − GC B cells from wild type litter mates ( bottom left ) and Igβ-Tg mice ( bottom right ). Numbers indicate the percentages of cells in each gate. ( c ) Serum anti-NP titers in each isotype after the immunization. Igβ-Tg (open circle) and wild type litter mates (closed circle) were immunized with NP-CGG on day 0 (1°) and boosted 42 d later (2°). Sera were collected at indicated day and assayed by ELISA as described in Methods section. Error bars indicate SEM for each group. Each group consists of 5 mice. Representative data from 3 repeated experiments is shown. ( d ) Affinity maturation of anti-NP IgG1 antibody in NP-CGG immunized Igβ-Tg (closed bar) and wild type litter mates (open bar). Error bars indicate SEM for each group. ( e ) Decreased frequency of somatic hypermutation in immunized Igβ-Tg mice. CD38 − GL7 + GC B cells were isolated from Igβ-Tg and wild type litter mates 21 days after the immunization. Total RNA was extracted and reverse transcribed to obtain cDNA samples. VH186.2 gene was then amplified and sequenced for the mutation analysis.

    Journal: Scientific Reports

    Article Title: Modulation of Igβ is essential for the B cell selection in germinal center

    doi: 10.1038/srep10303

    Figure Lengend Snippet: Forced expression of Igβ in B cells impaired the antibody production and affinity maturation. ( a ) Expression of Igβ in B220 + CD38 − GL7 − splenocytes from immunized Igβ-Tg (Igβ-Tg; line) and wild-type litter mates (wild type; shaded). ( b ) Expression of CD38 and GL7 in B220 + spleen cells from immunized wild type litter mates ( top left ) and Igβ-Tg mice ( top right ). Cell cycle analysis of CD38 − GL7 − GC B cells from wild type litter mates ( bottom left ) and Igβ-Tg mice ( bottom right ). Numbers indicate the percentages of cells in each gate. ( c ) Serum anti-NP titers in each isotype after the immunization. Igβ-Tg (open circle) and wild type litter mates (closed circle) were immunized with NP-CGG on day 0 (1°) and boosted 42 d later (2°). Sera were collected at indicated day and assayed by ELISA as described in Methods section. Error bars indicate SEM for each group. Each group consists of 5 mice. Representative data from 3 repeated experiments is shown. ( d ) Affinity maturation of anti-NP IgG1 antibody in NP-CGG immunized Igβ-Tg (closed bar) and wild type litter mates (open bar). Error bars indicate SEM for each group. ( e ) Decreased frequency of somatic hypermutation in immunized Igβ-Tg mice. CD38 − GL7 + GC B cells were isolated from Igβ-Tg and wild type litter mates 21 days after the immunization. Total RNA was extracted and reverse transcribed to obtain cDNA samples. VH186.2 gene was then amplified and sequenced for the mutation analysis.

    Article Snippet: Measurement of autoantibody and proteinuria Serum anti-dsDNA titers were measured using mouse anti-dsDNA IgG ELISA kit (Alpha Diagnostic International Inc.) according to the manufacturer’s instruction.

    Techniques: Expressing, Mouse Assay, Cell Cycle Assay, Enzyme-linked Immunosorbent Assay, Isolation, Amplification, Mutagenesis

    Detection sensitivity of anti- C. trachomatis antibodies in sera from Ctr-NAAT-positive women. An artificial population was created by combining 125 high anti- C. trachomatis antibody-prevalence sera from Ctr-NAAT-positive women ( 21 ) and 87 sera that had tested negative in multiple previous Ctr antibody assays ( Fig. 1 ). The first (leftmost) column indicates origin of sera from either the cohort of 125 Ctr NAAT-positive women (red) or the 3 cohorts of low Ctr-antibody-prevalence sera shown in Fig. 1 . The next column shows the ethnicity of the subjects as in Fig. 1 , followed by the composite reference standard CRS1 for NAAT-proven exposure to C. trachomatis ( Table 2 ). The next columns show background-corrected colorimetric signals of Ctr Mix1 and Ctr Mix 2 for an anti-human IgG polyclonal antibody conjugate, as well as monoclonal antibody conjugates for detection of short-lived IgG3 and IgA1 A2 antibody isotypes. The cutoff in these assays was chosen at 98.9% or higher specificity, with negative (white) and percentile reactivity ranking from low (light brown) to maximal (dark brown). The next columns indicate reactivity in 4 anti- C. trachomatis IgG commercial ELISAs as partially shown in Fig. 1 , followed by a Chlamydia genus-specific ELISA for detection of IgG antibodies against chlamydial LPS. The last column shows a consensus antibody status (positive/negative) based on both 4 commercial ELISAs and 2 peptide assays for anti- C. trachomatis IgG, IgG3, and IgA antibodies. This consensus status, CRS2, was used for ROC analyses that evaluated the performance of the individual assays ( Table 3 ).

    Journal: mSphere

    Article Title: Mixed Chlamydia trachomatis Peptide Antigens Provide a Specific and Sensitive Single-Well Colorimetric Enzyme-Linked Immunosorbent Assay for Detection of Human Anti-C. trachomatis Antibodies

    doi: 10.1128/mSphere.00484-18

    Figure Lengend Snippet: Detection sensitivity of anti- C. trachomatis antibodies in sera from Ctr-NAAT-positive women. An artificial population was created by combining 125 high anti- C. trachomatis antibody-prevalence sera from Ctr-NAAT-positive women ( 21 ) and 87 sera that had tested negative in multiple previous Ctr antibody assays ( Fig. 1 ). The first (leftmost) column indicates origin of sera from either the cohort of 125 Ctr NAAT-positive women (red) or the 3 cohorts of low Ctr-antibody-prevalence sera shown in Fig. 1 . The next column shows the ethnicity of the subjects as in Fig. 1 , followed by the composite reference standard CRS1 for NAAT-proven exposure to C. trachomatis ( Table 2 ). The next columns show background-corrected colorimetric signals of Ctr Mix1 and Ctr Mix 2 for an anti-human IgG polyclonal antibody conjugate, as well as monoclonal antibody conjugates for detection of short-lived IgG3 and IgA1 A2 antibody isotypes. The cutoff in these assays was chosen at 98.9% or higher specificity, with negative (white) and percentile reactivity ranking from low (light brown) to maximal (dark brown). The next columns indicate reactivity in 4 anti- C. trachomatis IgG commercial ELISAs as partially shown in Fig. 1 , followed by a Chlamydia genus-specific ELISA for detection of IgG antibodies against chlamydial LPS. The last column shows a consensus antibody status (positive/negative) based on both 4 commercial ELISAs and 2 peptide assays for anti- C. trachomatis IgG, IgG3, and IgA antibodies. This consensus status, CRS2, was used for ROC analyses that evaluated the performance of the individual assays ( Table 3 ).

    Article Snippet: We conclude that the higher sensitivity of the peptide assays described here is the result of the use of multiple highly reactive B-cell epitopes chosen from several immunodominant C. trachomatis proteins, while three of the commercial anti- C. trachomatis IgG ELISAs are exclusively based on OmpA antigens.

    Techniques: Enzyme-linked Immunosorbent Assay

    Comparative assay sensitivities of mixed and individual C. trachomatis peptide antigen ELISAs. The performance of the best Ctr Mix1 mixed peptide assay (12 peptides per microtiter well) was compared to the anti- C. trachomatis antibody status that had been previously determined by combining individual test results of 11 of these peptides, or by the single test result of the top-ranked OmpA peptide ( 21 ). The sera used were from 125 Ctr NAAT-positive women ( Fig. 2 ) and from 32 anti- C. trachomatis antibody-negative blood donors or low-risk women. The columns indicate first (leftmost) the cohort and ethnic origin of the sera, followed by background-corrected anti-IgG OD signals of Ctr Mix1 (peptides 1 to 12), the average RLU reactivity of 11 individually tested peptides (peptides 1 to 11), and the single RLU reactivity of the CtrOmpA_313-339 peptide. Similarly, the next columns show seroreactivities for the short-lived human IgG3 antibody isotype, followed by the IgA isotype. The last column shows a consensus of anti- C. trachomatis antibody status (positive/negative) based on all peptide assays. This consensus status, CRS3, was used for ROC performance analyses of individual assays ( Table 4 ).

    Journal: mSphere

    Article Title: Mixed Chlamydia trachomatis Peptide Antigens Provide a Specific and Sensitive Single-Well Colorimetric Enzyme-Linked Immunosorbent Assay for Detection of Human Anti-C. trachomatis Antibodies

    doi: 10.1128/mSphere.00484-18

    Figure Lengend Snippet: Comparative assay sensitivities of mixed and individual C. trachomatis peptide antigen ELISAs. The performance of the best Ctr Mix1 mixed peptide assay (12 peptides per microtiter well) was compared to the anti- C. trachomatis antibody status that had been previously determined by combining individual test results of 11 of these peptides, or by the single test result of the top-ranked OmpA peptide ( 21 ). The sera used were from 125 Ctr NAAT-positive women ( Fig. 2 ) and from 32 anti- C. trachomatis antibody-negative blood donors or low-risk women. The columns indicate first (leftmost) the cohort and ethnic origin of the sera, followed by background-corrected anti-IgG OD signals of Ctr Mix1 (peptides 1 to 12), the average RLU reactivity of 11 individually tested peptides (peptides 1 to 11), and the single RLU reactivity of the CtrOmpA_313-339 peptide. Similarly, the next columns show seroreactivities for the short-lived human IgG3 antibody isotype, followed by the IgA isotype. The last column shows a consensus of anti- C. trachomatis antibody status (positive/negative) based on all peptide assays. This consensus status, CRS3, was used for ROC performance analyses of individual assays ( Table 4 ).

    Article Snippet: We conclude that the higher sensitivity of the peptide assays described here is the result of the use of multiple highly reactive B-cell epitopes chosen from several immunodominant C. trachomatis proteins, while three of the commercial anti- C. trachomatis IgG ELISAs are exclusively based on OmpA antigens.

    Techniques:

    IgG reactivities of 203 blood donor sera with four commercial C. trachomatis ELISAs and two Ctr Mix1 and Ctr Mix2 peptide antigen mixtures. To identify a set of negative-control sera for anti- C. trachomatis antibodies, a panel of 203 sera was analyzed by 4 commercial colorimetric ELISAs and 2 highly sensitive chemiluminescent C. trachomatis - specific peptide antigen assays ( 21 ): (i) 18 healthy, low-risk women, (ii) 95 healthy women, and (iii) 90 healthy men. The first (leftmost) column indicates the origin of sera from either the cohorts of 95 female blood donors (dark green) or 90 male blood donors (medium green), or 18 low-risk women never diagnosed with C. trachomatis infection (light green). The next column shows the ethnic origin of the subjects: African American or black (dark blue), mixed race (medium blue), and Caucasian or white (light blue). The following 4 columns indicate percentile reactivity ranks of 4 commercial anti- C. trachomatis IgG ELISAs using the manufacturer-recommended cutoffs. Next, a consensus anti- C. trachomatis antibody status (positive/negative) is shown that assumed any serum as positive that was positive in any of the preceding 4 commercial C. trachomatis ELISAs. The next two columns show background corrected signals with Ctr Mix1 (peptides 1 to 12) and Ctr Mix 2 (peptides 13 to 24) for polyclonal antibody conjugate against human IgG, with reaction strength indicated by shading from white (nonreactive) to black (maximally reactive). The following column shows the average of relative light unit signals (RLU) for the Ctr Mix1 and Mix2 seroreactivities. The next column shows the binomial antibody status (positive or negative) based on average RLU signals of Ctr Mix1 and Mix2. The last column shows a final consensus of antibody status (positive/negative) based on the 4 commercial ELISAs and 2 peptide mix assays for anti- C. trachomatis IgG antibodies. The 87 consensus-negative sera were used as negative controls in the subsequent experiment ( Fig. 2 ).

    Journal: mSphere

    Article Title: Mixed Chlamydia trachomatis Peptide Antigens Provide a Specific and Sensitive Single-Well Colorimetric Enzyme-Linked Immunosorbent Assay for Detection of Human Anti-C. trachomatis Antibodies

    doi: 10.1128/mSphere.00484-18

    Figure Lengend Snippet: IgG reactivities of 203 blood donor sera with four commercial C. trachomatis ELISAs and two Ctr Mix1 and Ctr Mix2 peptide antigen mixtures. To identify a set of negative-control sera for anti- C. trachomatis antibodies, a panel of 203 sera was analyzed by 4 commercial colorimetric ELISAs and 2 highly sensitive chemiluminescent C. trachomatis - specific peptide antigen assays ( 21 ): (i) 18 healthy, low-risk women, (ii) 95 healthy women, and (iii) 90 healthy men. The first (leftmost) column indicates the origin of sera from either the cohorts of 95 female blood donors (dark green) or 90 male blood donors (medium green), or 18 low-risk women never diagnosed with C. trachomatis infection (light green). The next column shows the ethnic origin of the subjects: African American or black (dark blue), mixed race (medium blue), and Caucasian or white (light blue). The following 4 columns indicate percentile reactivity ranks of 4 commercial anti- C. trachomatis IgG ELISAs using the manufacturer-recommended cutoffs. Next, a consensus anti- C. trachomatis antibody status (positive/negative) is shown that assumed any serum as positive that was positive in any of the preceding 4 commercial C. trachomatis ELISAs. The next two columns show background corrected signals with Ctr Mix1 (peptides 1 to 12) and Ctr Mix 2 (peptides 13 to 24) for polyclonal antibody conjugate against human IgG, with reaction strength indicated by shading from white (nonreactive) to black (maximally reactive). The following column shows the average of relative light unit signals (RLU) for the Ctr Mix1 and Mix2 seroreactivities. The next column shows the binomial antibody status (positive or negative) based on average RLU signals of Ctr Mix1 and Mix2. The last column shows a final consensus of antibody status (positive/negative) based on the 4 commercial ELISAs and 2 peptide mix assays for anti- C. trachomatis IgG antibodies. The 87 consensus-negative sera were used as negative controls in the subsequent experiment ( Fig. 2 ).

    Article Snippet: We conclude that the higher sensitivity of the peptide assays described here is the result of the use of multiple highly reactive B-cell epitopes chosen from several immunodominant C. trachomatis proteins, while three of the commercial anti- C. trachomatis IgG ELISAs are exclusively based on OmpA antigens.

    Techniques: Negative Control, Infection

    FOXP1 overexpression specifically inhibits formation of IgG- but not IgM-secreting PCs. (A-B) CD19 + CD27 + MBCs were sorted from human peripheral blood and transduced with FOXP1-IRES-YFP, BCL6-IRES-GFP, or control empty vector and cultured under conditions that promote PC differentiation. Six days after transduction, YFP- or GFP-positive cells of cultures transduced with FOXP1, BCL6, or control vector were sorted. (A) Equal numbers of sorted cells (50 000) were cultured with IL-21 and IL-2 for an additional 24 hours. Thereafter, the supernatants were collected, and IgM and IgG protein levels were analyzed by ELISA. Levels were normalized to levels in control-transduced cells. Means ± SD of at least 4 independent experiments are shown. (B) Equal numbers of sorted cells were plated onto membranes in serial dilutions and cultured with IL-21 and IL-2 for an additional 18 hours, after which numbers of IgM- or IgG-secreting cells were determined by ELISPOT. Spot numbers were normalized to numbers in stimulated, control-transduced cells. Means ± SD of 3 independent experiments are shown (1 sample t test, * P

    Journal: Blood

    Article Title: The forkhead transcription factor FOXP1 represses human plasma cell differentiation

    doi: 10.1182/blood-2015-02-626176

    Figure Lengend Snippet: FOXP1 overexpression specifically inhibits formation of IgG- but not IgM-secreting PCs. (A-B) CD19 + CD27 + MBCs were sorted from human peripheral blood and transduced with FOXP1-IRES-YFP, BCL6-IRES-GFP, or control empty vector and cultured under conditions that promote PC differentiation. Six days after transduction, YFP- or GFP-positive cells of cultures transduced with FOXP1, BCL6, or control vector were sorted. (A) Equal numbers of sorted cells (50 000) were cultured with IL-21 and IL-2 for an additional 24 hours. Thereafter, the supernatants were collected, and IgM and IgG protein levels were analyzed by ELISA. Levels were normalized to levels in control-transduced cells. Means ± SD of at least 4 independent experiments are shown. (B) Equal numbers of sorted cells were plated onto membranes in serial dilutions and cultured with IL-21 and IL-2 for an additional 18 hours, after which numbers of IgM- or IgG-secreting cells were determined by ELISPOT. Spot numbers were normalized to numbers in stimulated, control-transduced cells. Means ± SD of 3 independent experiments are shown (1 sample t test, * P

    Article Snippet: IgG isotype ELISA was performed using the human IgG subclass profile ELISA kit (Invitrogen) according to the manufacturer’s instructions.

    Techniques: Over Expression, Transduction, Plasmid Preparation, Cell Culture, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot

    FOXP1 overexpression inhibits differentiation of IgG + MBCs but cannot further repress differentiation of IgM + MBCs. IgM + (IgG − IgA − ) and IgG + (IgM − IgA − ) MBCs were sorted from human peripheral blood and transduced with FOXP1-IRES-YFP, BCL6-IRES-GFP (except for IgG + MBCs), or control empty vector and cultured under conditions that promote PC differentiation. (A) Equal numbers of sorted cells (50 000) were cultured with IL-21 and IL-2 for an additional 24 hours. Thereafter, the supernatants were collected, and IgM and IgG protein levels were analyzed by ELISA. Levels were normalized to levels in control-transduced cells. Means ± SD of 3 independent experiments are shown. (B) Equal numbers of sorted cells were plated onto membranes in serial dilutions and cultured with IL-21 and IL-2 for an additional 18 hours, after which numbers of IgM- or IgG-secreting cells were determined by ELISPOT. Spot numbers were normalized to numbers in stimulated, control-transduced cells. Means ± SD of 3 independent experiments are shown (1 sample t test, * P

    Journal: Blood

    Article Title: The forkhead transcription factor FOXP1 represses human plasma cell differentiation

    doi: 10.1182/blood-2015-02-626176

    Figure Lengend Snippet: FOXP1 overexpression inhibits differentiation of IgG + MBCs but cannot further repress differentiation of IgM + MBCs. IgM + (IgG − IgA − ) and IgG + (IgM − IgA − ) MBCs were sorted from human peripheral blood and transduced with FOXP1-IRES-YFP, BCL6-IRES-GFP (except for IgG + MBCs), or control empty vector and cultured under conditions that promote PC differentiation. (A) Equal numbers of sorted cells (50 000) were cultured with IL-21 and IL-2 for an additional 24 hours. Thereafter, the supernatants were collected, and IgM and IgG protein levels were analyzed by ELISA. Levels were normalized to levels in control-transduced cells. Means ± SD of 3 independent experiments are shown. (B) Equal numbers of sorted cells were plated onto membranes in serial dilutions and cultured with IL-21 and IL-2 for an additional 18 hours, after which numbers of IgM- or IgG-secreting cells were determined by ELISPOT. Spot numbers were normalized to numbers in stimulated, control-transduced cells. Means ± SD of 3 independent experiments are shown (1 sample t test, * P

    Article Snippet: IgG isotype ELISA was performed using the human IgG subclass profile ELISA kit (Invitrogen) according to the manufacturer’s instructions.

    Techniques: Over Expression, Transduction, Plasmid Preparation, Cell Culture, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot